Two-dimensional zymogram analysis of nucleases in Bacillus subtilis
| Autor: | D. MacDonald Green, Susan A. Coughlin |
|---|---|
| Rok vydání: | 1983 |
| Předmět: |
Electrophoresis
Nuclease Deoxyribonucleases biology Isoelectric focusing Biophysics Sodium Dodecyl Sulfate Cell Biology Bacillus subtilis Hydrogen-Ion Concentration biology.organism_classification Biochemistry DNA-binding protein chemistry.chemical_compound Ribonucleases chemistry biology.protein Zymography Sodium dodecyl sulfate Isoelectric Focusing Deoxyribonuclease I Molecular Biology |
| Zdroj: | Analytical biochemistry. 133(2) |
| ISSN: | 0003-2697 |
| Popis: | A two-dimensional zymogram procedure for the analysis of nucleases is described. Isoelectric focusing (IEF) and nonequilibrium pH gradient electrophoresis (NEPHGE) were compared as first dimensions in combination with sodium dodecyl sulfate (SDS) electrophoresis as the second dimension in analyzing nucleases in lysates of Bacillus subtilis. All renaturable nucleases detected following SDS electrophoresis alone were resolved in NEPHGE-SDS electrophoresis gels whereas, in IEF gels, most either were at the basic end or were not present in the second-dimension gels. This method of analysis has revealed a complexity in nuclease species in B. subtilis not previously recognized. Eighty-three discreet nuclease activities have been detected in B. subtilis lysates. Using purified deoxyribonuclease I (bovine pancreas), as little as 10 pg of nuclease can be detected. |
| Databáze: | OpenAIRE |
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